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The mechanisms of inactivation against Gram‐negative bacteria (GNB) including Pseudomonas aeruginosa, Escherichia coli, and Vibrio parahaemolyticus, and Gram‐positive bacteria (GPB) involving Listeria monocytogenes and Staphylococcus aureus, by dielectric barrier discharge cold atmospheric plasma were elucidated. Both GNB and GPB were exposed to treatment with cold plasma produced from argon/oxygen mixture (90:10) as the working gas for different treatment times (1, 2.5, and 5 min). Survival counts were reduced with augmenting treatment time (TT; p < .05). All tested bacteria were not detected after TT of 5 min. There was an irreversible poration in cell wall of GNB. Meanwhile, GPB were partially shrunken after TT of 5 min. Therefore, irreversible cell wall damage, oxidation, and release of intracellular compounds including lipids, proteins and DNA were the proposed mode of action of cold …